Transient expression of human papillomavirus type 16 virus-like particles in tobacco and tomato using a tobacco rattle virus expression vector

0345055 - ÚEB 2011 RIV NL eng J - Journal Article
Šmídková, Markéta - Müller, M. - Thönes, N. - Piuko, K. - Angelisová, Pavla - Velemínský, Jiří - Angelis, Karel
Transient expression of human papillomavirus type 16 virus-like particles in tobacco and tomato using a tobacco rattle virus expression vector.
[Tranzientní exprese pseudovirionů lidského papilomaviru 16 v tabáku a rajčeti pomocí expresního vektoru odvozeného z Rattle viru tabáku.]
Biologia Plantarum. Roč. 54, č. 3 (2010), s. 451-460. ISSN 0006-3134. E-ISSN 1573-8264
R&D Projects: GA ČR GA521/01/1418; GA ČR GA521/04/0971; GA AV ČR IAA6038201; GA AV ČR IBS5038304; GA MŠMT 1M0505
Institutional research plan: CEZ:AV0Z50380511; CEZ:AV0Z50520514
Keywords : ELISA * Lycopersicon esculentum * Nicotiana benthamiana
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 1.582, year: 2010

The major capsid protein L1 of human papillomavirus type 16 (HPV16) was transiently expressed in tobacco (Nicotiana benthamiana and Nicotiana tabacum) and tomato (Lycopersicon esculentum) leaves using Agrobacterium tumefaciens. The expression vector pTV00 was derived from tobacco rattle virus (TRV). The highest L1 expression 15 mg g-1 was achieved when the coding sequence of L1 was optimized for expression in humans that caused an increase of the GC content from 38.2 % in wild type HPV16 to 64.1 % in optimized sequence. L1 monomers readily self-assembled into capsomeres and further into virus like particles (VLPs). Immunological characterization and electron microscopy showed that 89 % of L1 retained this structure also in extracts prepared from freeze-dried leaves. Plant expressed L1 was highly immunogenic without any additional adjuvant as vaccinated mice developed strong humoral and cellular immune response, comparable to that elicited by purified VLPs derived from insect cells.
Permanent Link: http://hdl.handle.net/11104/0186420